Bahig El-Deeb

Bacterial Load of Fresh Vegetables and Their Resistance to the Currently Used Antibiotics in Saudi Arabia

This study was carried out to describe the bacterial load and the occurrence of some disease-causing enteric bacteria on raw vegetables sold in Saudi markets. The study further aimed to analyze antibiotic resistance rates, production of extended-spectrum beta lactamase, and plasmid carriage among bacterial population of raw vegetables. Results revealed that none of them contained Bacillus cereus, Salmonella, and Escherichia coli O157:H7. However, Staphylococcus aureus and Shigella were detected in 11.8% and 4.4% of the samples, respectively. The bacterial loads ranged from 3 to 8 log(10) CFUg(-1) for aerobic bacteria and 1 to 4 log(10) CFUg(-1) for coliforms as well as Enterobacteriaceae. The isolates exhibited resistance in decreasing order for ampicillin (76.5%), cephalothin (69.5%), trimethoprime-sulfamethoxazole (36.7%), aminoglycosides (21.9%), tetracycline (17.2%), fluoroquinolones (17.2%), amoxycillin-clavulanic acid (13.3%), and chloramphenicol (7.8%). Maximum resistance to extended-spectrum beta-lactam antibiotics occurred in 14.8% of isolates and the production of extended-spectrum beta-lactamase was achieved by 2.3% of isolates. Multiple resistances to four or more antimicrobial agents along with plasmid with varied sizes were documented. These investigations indicate the occurrence of antibiotic resistance and plasmid carriage among bacterial isolates populating raw vegetables.

Isolation and characterization of endophytic bacteria from Plectranthus tenuiflorus medicinal plant in Saudi Arabia desert and their antimicrobial activities

Abstract The diversity and beneficial characteristics of endophytic microorganisms have been studied in Plectranthus tenuiflorus medicinal plant. However, information regarding naturally occurring P. tenuiflorus plant associated endophytes among different organs of host is limited. Endophytic bacteria were isolated from root, stem, and leaves of P. tenuiflorus plant. Among 28 endophytic bacterial isolates from different organs of P. tenuiflorus plant, 8 isolates were identified by partial sequencing of their 16S rRNA gene. The isolated endophytic bacteria were Bacillus sp., Bacillus megaterium, Bacillus pumilus, Bacillus licheniformis, Micrococcus luteus, Paenibacillus sp., Pseudomonas sp., and Acinetobacter calcoaceticus. The most isolates that exhibited extracellular enzymatic activity were belonged to the genus Bacillus. Furthermore, Bacillus sp. (HE613660) exhibited the stronger activities in extracellular enzymes such as amylase, esterase, lipase, protease, pectinase, xylanase, and cellulase than other strains. Considerable antimicrobial activities against a panel of human pathogenic microorganisms (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Streptococcus agalactiae, Proteus mirabilis, and Candida albicans) were recorded using crude extracts of the collected endophytic strains.

Molecular Screening of Streptomyces Isolates for Antifungal Activity and Family 19 Chitinase Enzymes

Thirty soil-isolates of Streptomyces were analyzed to determine their antagonism against plant-pathogenic fungi including Fusarium oxysporum, Pythium aristosporum, Colletotrichum gossypii, and Rhizoctonia solani. Seven isolates showed antifungal activity against one or more strain of the tested fungi. Based on the 16S rDNA sequence analysis, these isolates were identified as Streptomyces tendae (YH3), S. griseus (YH8), S. variabilis (YH21), S. endus (YH24), S. violaceusniger (YH27A), S. endus (YH27B), and S. griseus (YH27C). The identity percentages ranged from 98 to 100%. Although some isolates belonged to the same species, there were many differences in their cultural and morphological characteristics. Six isolates out of seven showed chitinase activity according to a chitinolytic activity test and on colloidal chitin agar plates. Based on the conserved regions among the family 19 chitinase genes of Streptomyces sp. two primers were used for detection of the chitinase (chiC) gene in the six isolates. A DNA fragment of 1.4 kb was observed only for the isolates YH8, YH27A, and YH27C. In conclusion, six Streptomyces strains with potential chitinolytic activity were identified from the local environment in Taif City, Saudi Arabia. Of these isolates, three belong to family 19 chitinases. To our knowledge, this is the first reported presence of a chiC gene in S. violaceusniger YH27A.

Characterization of endophytic bacteria associated with rose plant (Rosa damascena trigintipeta) during flowering stage and their plant growth promoting traits

Little is known about the bacterial communities associated with the rose plants inhabiting dry desert ecosystems. The aim of this study was to isolate and characterize endophytic bacteria from different organs of rose plant. Endophytic bacteria were observed in healthy roots, stems, leaves, and flowers of rose plant, with a significantly higher density in roots, followed by stems, leaves, and petals. A total of 38 bacterial endophytes were isolated and are closely related phylogenetically to Acetobacter, Acinetobacter, Methylococcus, Bacillus, Micrococcus, Planococcus by 16S rRNA sequence analysis. Six endophytic bacteria were found to produce IAA, solubilize Ca3(PO4)2 and produce siderophore. The six endophytic bacteria all had the capacity to produce hydrolytic enzyme such as cellulase, xylanase, pectinase, amylase, protease, lipase, and chitinase, but difference existed among these isolates.

Molecular Identification and Biofilm-Forming Ability of Culturable Aquatic Bacteria In Microbial Biofilms Formed in Drinking Water Distribution Networks

Drinking water distribution networks are known to harbor microbial biofilms. The aim of the present work is to (i) identify the culturable bacteria presented in the drinking-water distribution network, (ii) investigate the ability of isolated bacteria to form biofilm under some environmental stress conditions and some eliminating or removing treatments. To achieve it, 57 strains were isolated from biofilm (43 isolates) and water samples (14 isolates) collected from five stations in drinking-water distribution network in Taif city, Kingdom of Saudi Arabia (KSA). Partial sequences of 16S rRNA gene in the 57 isolates ensured the presence of only 22 different strains in biofilm samples. Among these strains, only 14 strains were also detected in water samples. Gram-negative Aeromonas hydrophila was the most occurred bacterium in the microbial biofilm obtained from the purified-water storage tanks followed by Gram-negative Pseudomonas sp. Gram-positive Bacillus subtilis was the most occurred bacterium in the microbial biofilm collected from the ends of the distribution pipes. Among the 22 isolated strains, 13 strains were strong biofilm producers at 30 and 37°C. The effects of environmental stresses including nutrient starvation (diluted TSB, 20:1), heating (100°C for 10 min), UV-treatment (240 nm for 10 min) and dynamic incubation (150 rpm min−1) on the formation of biofilm were also investigated. These conditions affected the biofilm formation ability of the isolated strains at different levels. Nutrient starvation enhanced biofilm formation by most of the isolates. Among some biofilm deforming treatments, SDS and trypsin had considerable effects on preventing biofilm formation by most of the isolated strains. In conclusion, the results of the present work indicated that not all biofilm strains released from biofilm to the drinking water. Also, not all biofilm strains were able to form biofilm. Most of isolated bacteria had ability to form biofilm at suboptimum temperature of growth. These results may provide basic information on formation of microbial biofilms and overcome the problem of deteriorating of water quality in the drinking-water distribution networks.

Molecular Characterization of Endophytic Bacteria from Metal Hyperaccumulator Aquatic Plant (Eichhornia crassipes) and Its Role in Heavy Metal Removal

In this study, among a collection of heavy metals resistant endophytic bacterial strains isolated from aquatic hyperaccumulator plant (Eichhornia crassipes), one plant growth promoting endophytic bacteria (PGPE), SVUB4 was selected for its ability to utilize 1-aminocyclopropane-1-carboxylic acid (ACC) as the sole N source and accumulate different heavy metals. The SVUB4 strain was characterized as Enterobacter sp. on the basis of its 16S rDNA sequences. Assessment of the parameters of plant growth promotion revealed the intrinsic ability of the strain for the production of IAA, siderophore and solubilization of insoluble phosphate. Furthermore, plasmid DNA analysis of Enterobacter sp. strain SVUB4 indicated the presence of a single large plasmid element. The results of plasmid curing experiments demonstrated that the ability of this strain to grow in presence of Cd and Zn was encoded by the 98 kb plasmid, whereas the ability to grow in the presence of Pb appeared to be encoded by the chromosome. The Cd and Zn removal capacity of the respective metal sensitive strain (plasmidless) were about 36 and 45 μg/g-1 DW, respectively, while the removal capacity of the both metal by metal resistant strain (p SVUB4) showed a significantly higher Cd and Zn removal capacity of 153 and 228 μg/g−1 DW, respectively. However, both strains exhibited a similar pattern of Pb accumulation. The present observation also showed that for wild-type strain SVUB4 (pSVUB4), the overall level of IAA production in the absence and in the presence of Cd2+ or Zn2+was approximately the same. Nevertheless, strain SVUB4M in this respect appeared to be more sensitive to heavy metals: a noticeable decrease in IAA production was observed under the effect of both metals, especially with Cd2+.

Isolation and Characterization of Endophytic Bacilli Bacterium from Maize Grains Able to Detoxify Aflatoxin B1

Aflatoxins are commonly found in cereals worldwide and bring significant threats to the food industry and animal food production. Among a collection of aflatoxin-degradative endophytic bacteria isolated from grains of corn plant, the strain TUBF1 was selected based on its high-ability to utilize aflatoxin B1 (AFB1) (10 μg/mL) as the sole carbon source. The morphological, physiological, and phylogenetic studies indicated that strain TUBF1 belonged to the Bacillus sp. group. HPLC was used to determine the reduction in AFB1 concentrations. Bacillus sp. TUBF1 had the strongest ability to detoxify toxin, where the degradation percentages of AFB1 was 81.5% and 100% after 48 and 72 h, respectively. The degradation of AFB1 was mainly in the culture supernatant of TUBF1 rather than its cells. About 90% of AFB1 was degraded within the first 12 h and reduced to the undetectable level after 24 h. The supernatant was characterized by considerable activity at wide range of temperatures (10–40°C) with optimal activity at 32°C and pH 6.5–7.5. Biosafety assessment test indicated that the crude enzymes had the high ability to detoxify AFB1. In conclusion, this is the first report on AFB1 detoxification using an endophytic Bacillus isolated from grains.

Mycobiota of Oil-Contaminated Soil Samples and Their Abilities for Dibenzothiophene Desulfurization

ABSTRACT High sulfur content of crude oil leads to poor quality of oil products and many other negative consequences such as corrosion, catalyst poisoning and environmental pollution. Saudi Arabia is seeking to reduce sulfur content in diesel and gasoline to 10 ppm and to lower benzene content in gasoline to 1%. Biotechnological processes such as biodesulfurization can be considered an alternative or complement to conventional oil refining technologies. So, the objective of the present project is to isolate and identify endogenous fungal isolates capable of oil biodesulfurization. From 60 oil-contaminated soil samples collected from Saudi Arabia, 15 species belonged to 9 fungal genera were collected and identified morphologically and with ITS sequencing. Members of Aspergillus, Penicillium and Fusarium were the most prevalent in the investigated samples. Among the collected fungal species, only Stachybotrys bisbyiisolates were able to utilize dibenzothiophene (DBT) as the sole sulfur source. Stachybotrys bisbyi TUSb1 could desulfurize 99% of the DBT (0.3 mM) as the sulfur source by a co-metabolism reaction with other carbon sources through the same pathway as 4S (involves sequential oxidation of the sulfur part and cleaving of the C–S bonds), and produced 2-hydroxy biphenyl (2-HBP) during 7 days of incubation at 30°C and 180 rpm. Stachybotrys bisbyi TUSb1 showed broad specificity for removing sulfur in different sulfur-containing compounds.

Antibiofilm potential of biogenic silver nanoparticles against Kocuria rosea And Kocuria rhizophila

The present study aims at biosynthesizing, characterizing and evaluating the biogenic silver nanoparticles (AgNPs) as antimicrobial and antibiofilm against Kocuria rosea and Kocuria rhizophila. Cellfree supernatant of Proteus mirabilis culture was used for biosynthesizing AgNPs, which confirmed by visualizing color change and X-ray diffraction. Transmission electron microscopy showed the formation of AgNPs in the range of 5–40 nm. ART-FTIR spectra provided evidence for presence of proteins as possible biomolecules responsible for stability of AgNPs and act as capping agent. AgNPs had ability to inhibit growth of K. rosea and K. rhizophila. The minimum inhibitory concentration (MIC90) of AgNPs against both strains was 25 μg/mL. Antiadhesive effect of AgNPs was verified at sub-MIC90 dose (12.5 μg/mL). The AgNPs concentrations up to 100 μg/mL were not effective for complete removing the already established biofilms with maximum removing percentage of 30.5–34.9%. In conclusion, the present study demonstrated an unprecedented green process for biosynthesizing stable spherical-shaped AgNPs. Early control is suggested by preventing biofilm formation using low AgNPs concentration (12.5 μg/mL) as a potential ingredient for formulating effective chemical sanitizers.

Plasmid-mediated Molluscicide Bayluscide Degradation by Bacterial Strain Isolated from Schistosome Vector Snail Bulinus truncates

Pseudomonas spp. strain Bal1 was isolated from Schistosome vector snails Bulinus Truncates. Strain Bal1 was identified as Pseudomonas putida using partial sequence of 16s rRNA. This strain was able to utilize a Bayluscide as a sole source of carbon and nitrogen. The degradation of Bayluscide by Bal1 strain is mediated by pBal1 (110 Kb) plasmid. The loss of the plasmid resulted irreversibly in a derivative strain that was unable to degrade Bayluscide. The transfer of these plasmids from wild-type strain Bal1 to Bal1M derivative restored completely its capability to degrade the molluscicide. It is proposed that pBal1 is a conjugative plasmid and is involved in the Bayluscide degradation. The effect of bacterial degradation upon toxicity was tested, and it was shown that the molluscicidal action of Bayluscide was significantly reduced by bacterial action.